期刊
ANALYTICAL CHEMISTRY
卷 83, 期 16, 页码 6196-6201出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac2016213
关键词
-
资金
- Health Initiative of the National Research Council of Canada
- Natural Sciences and Engineering Research Council of Canada
MicroRNAs (miRNAs) are small (similar to 22 nt) regulatory RNAs that are frequently deregulated in cancer and have shown promise as tissue- and blood-based biomarkers for cancer classification and prognostication. Here we present a protein-facilitated affinity capillary electrophoresis (ProFACE) assay for rapid quantification of miRNA levels in blood serum using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. The method utilizes either the selective binding of SSB to a single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. For the detection of ultralow amounts of miRNA without polymerase chain reaction (PCR) amplification in blood samples we apply off-line preconcentration of synthetic miRNA-122 from serum by p19-coated magnetic beads followed by online sample stacking in the ProFACE assay. The detection limit is 0.5 fM or 30 000 miRNA molecules in 1 mL of serum as a potential source of naive miRNAs.
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